Otimização da produção de amilases e pectinases por Gongronella butleri usando subprodutos agroindustriais
Carregando...
Data
2021-09-02
Autores
Braga, Héberly Fernandes
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Objetivou-se determinar e otimizar a produção de amilases e pectinases, por Gongronella butleri sob cultivo em estado sólido (CES), em subprodutos agroindustriais (SA). Foi realizado teste de verificação lítica a 35ºC, em meio líquido e sólido com amido ou pectina; assim como o crescimento micelial em ágar padrão a 25, 30, 35 e 40ºC. A produção enzimática foi obtida pelo CES a 25, 30 e 35ºC em 72 h, sobre diferentes SA caracterizados físico-quimicamente; e as atividades determinadas pelo método de redução do ácido-3,5-dinitrosalicílico (DNS), além do dextrinizante para as amilases. A temperatura foi fixada a 35ºC, e o perfil enzimático pelo DNS foi monitorado para alguns dos SA ao longo de 120 h, visando identificar os picos de maior atividade, que ocorreram em farelo de trigo (FT) e soja (FS). O acréscimo das atividades enzimáticas nesses SA foi testado por planejamento experimental 23, a 35ºC por 72 h, combinando proporções de FT e FS, e variando o teor inicial de solução mineral e CaCl2 dos cultivos; seguido por planejamento um fator por vez, onde se monitorou o efeito de combinações de FT e FS durante 120 h. As melhores condições experimentais obtidas anteriormente foram usadas como parâmetro para se aplicar um delineamento composto central rotacional (DCCR), visando obter o ponto ótimo de produção enzimática, tendo como fatores umidade inicial do meio; temperatura e tempo de cultivo. O fungo desenvolveu-se em meio líquido e sólido contendo amido ou pectina, formando massa micelial em até 24 h. A expansão micelial apresentou diferença significativa (p ≤ 0,05) inicial nas temperaturas testadas, após 72 h, não sendo observado crescimento a 40ºC. Dos SA avaliados, 88,9% induziram atividade amilolítica dextrinizante inferior a 2,5 U mL-1 em ao menos uma das temperaturas, sendo a maior atividade (9,6 U mL-1) observada a 35ºC em FT. As atividades amilolíticas pelo DNS foram ≥ 7,0 U g-1, em todas temperaturas testadas, quando se empregou FS e FT, e para este houve aumento de 31% da atividade pectinolítica de 25ºC para 35ºC. O pico de atividade amilolítica pelo DNS foi atingido em 72 h para FT (13,7 U g-1) e 96 h para FS (16,0 U g-1); os outros SA mantiveram perfis de atividade inferiores a 3,8 U g-1. A atividade pectinolítica atingiu pico de 41,7 U g-1 para FT, e 33,0 U g-1 para FS em 48 h e após emprego do planejamento 23 foi observado acréscimo de 82% na atividade amilolítica e 33% na pectinolítica, quando comparado à média individualizada em FT e FS. As maiores atividades amilolíticas (15,97 U g-1) e pectinolíticas (128,92 U g-1), tenderam a ocorrer em 48 h, e na combinação FT:FS (25%:75%), não diferindo de FT:FS (50%:50%), para pectinases, após planejamento um fator por vez. A ótima produção enzimática (amilases = 32,35 U g-1; pectinases = 246,62 U g-1) obtida por DCCR foi atingida em 61,37% umidade inicial, 30ºC e 68 h de cultivo. Os resultados indicam potencial de produção enzimática, em especial pectinases, por G. butleri quando em FT e FS.
The objective was to determine and optimize the production of amylases and pectinases by Gongronella butleri under solid state cultivation (SSC) in agro-industrial by-products (AP). A lytic verification test was carried out at 35ºC, in liquid and solid medium with starch or pectin; as well as mycelial growth on standard agar at 25, 30, 35 and 40ºC. The enzymatic production was obtained by SSC at 25, 30 and 35ºC in 72 h, on different physicochemically characterized AP; and the activities determined by the method of reduction of 3,5-dinitrosalicylic acid (DNS), in addition to the dextrinizer for amylases. The temperature was fixed at 35ºC, and the enzymatic profile by DNS was monitored for some of the AP over 120 h, in order to identify the peaks of higher activity, which occurred in wheat bran (WB) and soybean (SB). The addition of enzymatic activities in these AP was tested by experimental design 23, at 35ºC for 72 h, combining proportions of WB and SB, and varying the initial content of mineral solution and CaCl2 of the crops; followed by planning one factor at a time, where the effect of combinations of WB and SB was monitored during 120 h. The best experimental conditions previously obtained were used as a parameter to apply a central composite rotational design (DCCR), in order to obtain the optimum point of enzyme production, having as factors the initial moisture of the medium; temperature and time of cultivation. The fungus developed in a liquid and solid medium containing starch or pectin, forming mycelial mass within 24 h. The mycelial expansion showed a significant difference (p ≤ 0.05) at the initial temperatures tested, after 72 h, with no growth being observed at 40ºC. Of the AP evaluated, 88.9% induced dextrinizing amylolytic activity less than 2.5 U mL-1 in at least one of the temperatures, with the highest activity (9.6 U mL-1) being observed at 35ºC in WB. The amylolytic activities by DNS were ≥ 7.0 U g-1, at all temperatures tested, when SB and WB were used, and for this there was an increase of 31% in pectinolytic activity from 25ºC to 35ºC. The peak of amylolytic activity by DNS was reached in 72 h for WB (13.7 U g-1) and 96 h for SB (16.0 U g-1); the other AP maintained activity profiles below 3.8 U g-1. The pectinolytic activity reached a peak of 41.7 U g-1 for WB, and 33.0 U g-1 for SB in 48 h and after the use of planning 23, an increase of 82% in the amylolytic activity and 33% in the pectinolytic activity was observed, when compared to the individualized average in WB and SB. The highest amylolytic (15.97 U g-1) and pectinolytic (128.92 U g-1) activities tended to occur within 48 h, and in the WB:SB (25%:75%) combination, not differing from WB:SB (50%:50%), for pectinases, after planning one factor at a time. The optimal enzyme production (amylases = 32.35 U g-1; pectinases = 246.62 U g-1) obtained by DCCR was reached at 61.37% initial moisture, 30ºC and 68 h of cultivation. The results indicate potential for enzyme production, especially pectinases, by G. butleri when in WB and SB.
The objective was to determine and optimize the production of amylases and pectinases by Gongronella butleri under solid state cultivation (SSC) in agro-industrial by-products (AP). A lytic verification test was carried out at 35ºC, in liquid and solid medium with starch or pectin; as well as mycelial growth on standard agar at 25, 30, 35 and 40ºC. The enzymatic production was obtained by SSC at 25, 30 and 35ºC in 72 h, on different physicochemically characterized AP; and the activities determined by the method of reduction of 3,5-dinitrosalicylic acid (DNS), in addition to the dextrinizer for amylases. The temperature was fixed at 35ºC, and the enzymatic profile by DNS was monitored for some of the AP over 120 h, in order to identify the peaks of higher activity, which occurred in wheat bran (WB) and soybean (SB). The addition of enzymatic activities in these AP was tested by experimental design 23, at 35ºC for 72 h, combining proportions of WB and SB, and varying the initial content of mineral solution and CaCl2 of the crops; followed by planning one factor at a time, where the effect of combinations of WB and SB was monitored during 120 h. The best experimental conditions previously obtained were used as a parameter to apply a central composite rotational design (DCCR), in order to obtain the optimum point of enzyme production, having as factors the initial moisture of the medium; temperature and time of cultivation. The fungus developed in a liquid and solid medium containing starch or pectin, forming mycelial mass within 24 h. The mycelial expansion showed a significant difference (p ≤ 0.05) at the initial temperatures tested, after 72 h, with no growth being observed at 40ºC. Of the AP evaluated, 88.9% induced dextrinizing amylolytic activity less than 2.5 U mL-1 in at least one of the temperatures, with the highest activity (9.6 U mL-1) being observed at 35ºC in WB. The amylolytic activities by DNS were ≥ 7.0 U g-1, at all temperatures tested, when SB and WB were used, and for this there was an increase of 31% in pectinolytic activity from 25ºC to 35ºC. The peak of amylolytic activity by DNS was reached in 72 h for WB (13.7 U g-1) and 96 h for SB (16.0 U g-1); the other AP maintained activity profiles below 3.8 U g-1. The pectinolytic activity reached a peak of 41.7 U g-1 for WB, and 33.0 U g-1 for SB in 48 h and after the use of planning 23, an increase of 82% in the amylolytic activity and 33% in the pectinolytic activity was observed, when compared to the individualized average in WB and SB. The highest amylolytic (15.97 U g-1) and pectinolytic (128.92 U g-1) activities tended to occur within 48 h, and in the WB:SB (25%:75%) combination, not differing from WB:SB (50%:50%), for pectinases, after planning one factor at a time. The optimal enzyme production (amylases = 32.35 U g-1; pectinases = 246.62 U g-1) obtained by DCCR was reached at 61.37% initial moisture, 30ºC and 68 h of cultivation. The results indicate potential for enzyme production, especially pectinases, by G. butleri when in WB and SB.
Descrição
Palavras-chave
Biomassa vegetal, Bioprocesso, Fungos isolados do cerrado, Produção enzimática, Resíduos agroindustriais, Vegetable biomass, Bioprocess, Fungi isolated from the cerrado, Enzymatic production, Agro-industrial waste